Mechanism of Xiaoer Chiqiao Qingre Granules in clearing heat and removing food stagnation in suckling rats with fever and food accumulation based on metabolomics / 中国中药杂志

Children's fever is often accompanied by food accumulation. Traditional Chinese medicine believes that removing food stagnation while clearing heat of children can effectively avoid heat damage. To systematically evaluate the efficacy of Xiaoer Chiqiao Qingre Granules(XRCQ) in clearing heat and removing food accumulation and explore its potential mechanism, this study combined suckling SD rats fed with high-sugar and high-fat diet with injection of carrageenan to induce rat model of fever and food accumulation. This study provided references for the study on the pharmacodynamics and mechanism of XRCQ. The results showed that XRCQ effectively reduced the rectal temperature of suckling rats, improved the inflammatory environment such as the content of interleukin-1β(IL-1β), interleukin-2(IL-2), interferon-γ(IFN-γ), white blood cells, and monocytes. XRCQ also effectively repaired intestinal injury and enhanced intestinal propulsion function. According to the confirmation of its efficacy of clearing heat, the thermolytic mechanism of XRCQ was further explored by non-targeted and targeted metabolomics methods based on LTQ-Orbitrap MS/MS and UPLC-QQQ-MS/MS. Non-target metabolomics analysis of brain tissue samples was performed by QI software combined with SIMCA-P software, and 22 endogenous metabolites that could be significantly regulated were screened out. MetaboAnalyst pathway enrichment results showed that the intervention mechanism was mainly focused on tyrosine metabolism, tricarboxylic acid cycle, inositol phosphate metabolism, and other pathways. At the same time, the results of targeted metabolomics of brain tissue samples showed that XRCQ changed the vitality of digestive system, and inhibited abnormal energy metabolism and inflammatory response, playing a role in clearing heat and removing food stagnation from multiple levels.

A case report of polyglandular syndrome induced by programmed death-1 inhibitor and literature review / 中华内分泌代谢杂志

We reported a case of polyglandular syndrome induced by programmed death-1(PD-1) inhibitors. The patient was a 51-years-old male with non-small cell lung cancer, treated with PD-1 inhibitor nivolumab/pembrolizumab because of postoperative subcarinal lymph node metastasis indicated by PET-CT. During 14 cycles of PD-1 inhibitor treatment, the patient successively developed primary hypothyroidism, and type 1 diabetes mellitus(T1DM). More than five months after the withdrawal of pembrolizumab, the patient experienced recurrentce. Laboratory examinations showed mild hyponatremia and hypopituitarism including ACTH and growth hormone(GH)/insulin-like growth factor-1(IGF-1) insufficiency. This is the first report of a patient diagnosed as polyglandular syndrome caused by PD-1 inhibitor. In particularly, the hypothyroidism and T1DM did not improve after drug withdrawal, while hypopituitarism was further aggravated. This case reminds us that we should pay more attention to the changes of endocrine function during and after the treatment of PD-1 inhibitor, so that we can make the correct diagnosis and take proper medical measures timely, to avoide missed diagnosis, and improper treatment.

Analysis of clinical trials of dermatological drugs in China, 2020 / 中华皮肤科杂志

Objective:To deliver understanding of the latest research progress on clinical trials and approval of dermatological drugs in China in 2020.Methods:A registration and information disclosure platform for drug clinical studies and a query system for domestic and imported drugs in the National Medical Products Administration of China were searched for registered clinical trials and approved dermatological drugs, respectively. The number and stages of clinical trials, indications and classification of involved products, and listed dermatological drugs in 2020 were summarized and depicted.Results:There were 157 dermatological drug trials registered in China in 2020, accounting for 6.16% of all the 2 548 clinical drug trials, including 127 (80.9%) initiated by Chinese pharmaceutical enterprises and 25 (15.9%) international multicenter trials. Among the 127 drug trials initiated by Chinese pharmaceutical enterprises, bioequivalence trials were mostly common, accounting for 55.9% (71/127) . Compared with global pharmaceutical enterprises, domestic pharmaceutical companies initiated significantly decreased proportions of international multicenter trials (1.9% [3/157] vs. 14.0% [22/157], P < 0.001) , but significantly increased proportions of phaseⅠclinical trials and bioequivalence trials (24.4% [31/127] vs. 10.0% [3/30], 55.9% [71/127] vs. 0, respectively, both P < 0.001) . Totally, 90 kinds of dermatological drug were involved in all the trials, psoriasis, atopic dermatitis and melanoma were the most common indications, and innovative drugs accounted for 53.3% (48/90) ; the proportion of innovative drugs was significantly lower in domestic pharmaceutical companies than in global pharmaceutical companies (43.2% [32/74] vs. 16/16, P < 0.001) . In addition, 28 dermatological drugs developed by 22 pharmaceutical companies were approved in China in 2020, of which 21 drugs were developed by domestic pharmaceutical companies. Conclusion:Clinical drug trials carried out by domestic pharmaceutical companies mostly focus on generic drugs, and it is still necessary for domestic pharmaceutical companies to further improve the innovation ability.

Identification of chemical constituents of Xiaoer Chiqiao Qingre Granules based on UHPLC-LTQ-Orbitrap-MS/MS and network pharmacology analysis / 中国中药杂志

This study aimed to qualitatively analyze the chemical components in Xiaoer Chiqiao Qingre Granules(XRCQ) by UHPLC-LTQ-Orbitrap-MS/MS and identify its material basis. The absorbed components in plasma were combined for exploring the potential action mechanism by integrated network pharmacology. ACQUITY UPLC HSS T3(2.1 mm×100 mm, 1.8 μm) column and mobile phase system of 0.1% formic acid solution(A)-acetonitrile(B) were used for gradient elution, followed by high resolution liquid chromatography-mass spectrometry in both positive and negative ion scanning modes. According to the precise relative molecular mass and MS/MS fragment ions, a total of 124 chemical components were identified in XRCQ by the comparison with references and literature reports, among which 29 compounds were completely confirmed by comparison with reference substances. Then, the main absorbed components of XRCQ in plasma were also analyzed and clarified by UHPLC-LTQ-Orbitrap-MS/MS. BATMAN-TCM and SwissTargetPrediction were used for target prediction of absorbed components in plasma. Following the plotting of association network with Cytoscape 3.8.2, the core targets were subjected to GO and KEGG pathway enrichment analysis and a component-target-pathway network was constructed. A total of eight main targets of XRCQ against fever in children were obtained together with eight absorbed components in plasma, including glycyrhydinic acid, hesperidin, emodin, reticuline, daidzein, magnolignan C, magnolignan A, and magnolaldehyde D. It was inferred that XRCQ might improve alimentary system abnormality, inflammatory response, oxidative stress, and endocrine disorder through tumor necrosis factor, PI3 K-AKT, and other signaling pathways. The present study comprehensively expounded the chemical profiles of XRCQ and the main absorbed components in plasma and predicted the potential mechanism of XRCQ based on integrated network pharmacology, which has provided certain theoretical reference for the clinical application of XRCQ.

Identification of Bupleurum chinense and B. longiradiatum based on ITS2 barcode / 中国中药杂志

In this study, ITS2 barcode was used to identify Bupleurum chinense and B. longiradiatum. The ITS2 regions of 48 samples were amplified and sequenced. The sequences obtained above were aligned and the K2P distances were calculated. We used three methods, BLAST1, nearest distance and phylogenetic tree (NJ-tree), to test the identification ability. The results showed that the maximum intraspecific genetic distance of B. chinense was 0.013, and the minimum interspecific genetic distance between B. chinense and B. longiradiatum was 0.049. The NJ-tree can easily identify B. chinense and B. longiradiatum. Therefore, the ITS2 barcode is suitable to identify B. chinense and B. longiradiatum.

Molecular identification of Manis pentadactyla using DNA barcoding / 中国中药杂志

The COI gene as DNA barcode was used to identify the Manis pentadactyla and its adulterants in order to provide a scientific basis for the molecular identification of M. pentadactyla. Genomic DNA was extracted from experimental samples using the DNA extraction kit. The COI genes were amplified using polymerase chain reaction (PCR) and sequenced bi-directionally. Obtained sequences were assembled using the CodonCode Aligner. The neighbor-joining (NJ) tree was constructed by MEGA 6.0. The results indicated that COI sequences were successfully amplified and NJ trees results indicated that M. pentadactyla and its adulterants can be easily identification. Therefore, the COI gene is an efficient barcode for identification of M. pentadactyla and its adulterants,which will provide a new technique for the market supervision.

Effect of Colquhounia root tablet on IL-2 and IFN-γ mRNA expression in rats with experimental allergic encephalomyelitis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of Colquhounia root tablet on IL-2 and IFN-γ mRNA expression in experimental allergic encephalomyelitis (EAE) of rats.</p><p><b>METHODS</b>The allergic encephalomyelitis model was established in Wistar rats by immunization with myelin basic protein of spinal cord of guinea pig and complete Freund's adjuvant. The rats in treatment group received Colquhounia root tablet (300 mg*kg(-1), BID). The symptom of EAE was observed; pathological feature and myelin of brain and spinal cord were detected with HE stain and Loyez's stain, respectively. The expressions of IL-2 and IFN-γ mRNA were assayed by RT-PCR.</p><p><b>RESULTS</b>No EAE symptoms were developed in treatment group, the expressions of IL-2 and IFN-γ mRNA were 0.345 ± 0.032 and 0.353 ± 0.023, which were significantly lower than those of model group (P<0.01). The histopathologic examinations revealed that less inflammation cells around vessels and demyelination in white matter of brain and spinal cords were observed in treatment group than in model group.</p><p><b>CONCLUSION</b>Colquhounia root tablets are effective in treatment of EAE of rats, which may be associated with inhibition of the expression of IL-2 and IFN-γ mRNA.</p>

Expression of L - selectin in experimental allergic encephalomyelitis of Wistar rats and its clinical significance / 中国基层医药

Objective To investigate the effect of L-selectin in experimental allergic encephalomyelitis (EAE) of Wistar rats.Methods The rats were randomly divided into four groups;the normal group,the CFA group, the LMS group and the model group;The animal models were established in rats by immunization with myelin basic protein of spinal cord of guinea pig and complete Freund's adjuvant(CFA).The symptom of EAE was observed; pathological feature and myelin of brain and spinal were detected with HE stain and Loyez's stain respectively.The number of positive vessels of L-selectin expression was detected by immunohistochemistry.Results 100% experimental Wistar rats treated with MBP and levamisole developed EAE,but none of the other groups.The number of positive vessels of L-selectin expression was (31.86 ± 1.39) in model group, obviously higher than that of in the normal group (1.38 ±0.18) ,the CFA group( 1.50 ±0.27) and the LMS group(7.25 ±0.59) (all P <0.05) ;The inflammation cells were found around vessels and demyelination were seen in white matter of brain and spinal cords.Conclusion The expression of L-selectin should play an important role in EAE.

Influence of morning blood pressure on carotid atherosclerosis in elderly patients with essential hypertension / 中国基层医药

Objective To investigate the association between morniing blood pressure surge (MBPS) and carotid atherosclerosis in elder patients with essential hypertension. Methods According to the results of24h ambulatory blood pressure monitoring, 106 patients were classified as the morning BP surge group (MBPS group,n = 58) ,and nonsurge group (NMBPS group, n = 48). Patients underwent carotid ultrasound and the intima-medial thickness (CCA-IMT) and plaques were examined. Results The CCA-IMT of the MBPS group was significantly thicker than that the NMBPS group[(1.27 ± 0. 12)mm vs (0.92 ± 0.33 )mm], P ＜ 0. 05 ) ;②Compared with the NMBPS group,the severity of carotid arteries plaque of the MBPS group was significantly higher (72. 15% vs 54.21% ), ( P ＜0. 01 ) ;③Pearson relation analysis showed CCA-IMT level positively correlated with age (r = 0.288, P ＜ 0.001 ) ,the average of 24h SBP ( r = 0. 768 ,P ＜ 0. 001 ), and MBPS ( r = 0. 768, P ＜ 0.001 ). Conclusion The study showed that MBPS was closely related with carotid atherosclerosis in elder patients with essential hypertension and was an important risk factor in the process of atheresclerosis.

The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The genome of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) includes sequences encoding the putative protein X4 (ORF8, ORF7a), consisting of 122 amino acids. The deduced sequence contains a probable cleaved signal peptide sequence and a C-terminal transmembrane helix, indicating that protein X4 is likely to be a type I membrane protein. This study was conducted to demonstrate whether the protein X4 was expressed and its essential function in the process of SARS-CoV infection.</p><p><b>METHODS</b>The prokaryotic and eukaryotic protein X4-expressing plasmids were constructed. Recombinant soluble protein X4 was purified from E. coli using ion exchange chromatography, and the preparation was injected into chicken for rising specific polyclonal antibodies. The expression of protein X4 in SARS-CoV-infected Vero E6 cells and lung tissues from patients with SARS was performed using immunofluorescence assay and immunohistochemistry technique. The preliminary function of protein X4 was evaluated by treatment with and over-expression of protein X4 in cell lines. Western blot was employed to evaluate the expression of protein X4 in SARS-CoV particles.</p><p><b>RESULTS</b>We expressed and purified soluble recombinant protein X4 from E.coli, and generated specific antibodies against protein X4. Western blot proved that the protein X4 was not assembled in the SARS-CoV particles. Indirect immunofluorescence assays revealed that the expression of protein X4 was detected at 8 hours after infection in SARS-CoV-infected Vero E6 cells. It was also detected in the lung tissues from patients with SARS. Treatment with and overexpression of protein X4 inhibited the growth of Balb/c 3T3 cells as determined by cell counting and MTT assays.</p><p><b>CONCLUSION</b>The results provide the evidence of protein X4 expression following SARS-CoV infection, and may facilitate further investigation of the immunopathological mechanism of SARS.</p>

Preparation, identification, and analysis on tissue chips of polyclonal anti-peptide antibody to chemokine-like factor 1 / 中国医学科学院学报

<p><b>OBJECTIVE</b>To prepare the polyclonal anti-peptide antibody against chemokine-like factor1 (CKLF1) and apply it to the expression and functional studies of CKLF1.</p><p><b>METHODS</b>CKLF1 was analyzed with bioinformatics methods. The 16 amino acids sequence peptide was selected from CKLF1 C terminal end. Antibody was raised by immunizing rabbits with the peptide conjugated to keyhole limpet hemocyanin (KLH).</p><p><b>RESULTS</b>A high titer polycolonal antibody was obtained from the rabbit against the peptide. ELISA analysis proved that the titer of rabbit serum against anti-peptide of CKLF1 was up to 10(-4). Western blot analysis revealed that it could react not only with recombinant CKLF1 expressed in a cell-Free Protein Biosynthesis System and Drosophila S2 cells, but also recognize the endogenous CKLFs in the tissue array. Positive staining was detected in the normal bronchial cartilage, gastric mucosa, and gastric smooth muscle tissues. Normal rectum and well-differentiated rectal carcinoma showed strong positive staining, but the poor-differentiated rectal carcinoma samples revealed negative staining.</p><p><b>CONCLUSION</b>The anti-peptide antibody can specifically recognize CKLFs and may be a useful reagent for the detection of CKLF1.</p>

Efficacy observation in treating patients with postmenopausal coronary heart disease by bushen peiyuan principle / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Bushen Peiyuan Principle (BSPY, a TCM principle for tonifying Kidney and nourishing primordial energy) in treating patients with postmenopausal coronary heart disease (PCHD) instead of hormone treatment.</p><p><b>METHODS</b>Twenty-five healthy women, who were monepaused for over 5 years but without CHD complication were allocated in Group A, 25 patients with PCHD complication suffered from estrogen contraindications such as embolism, hysteromyoma and mammary adenoma, were arranged in Group B, and 25 patients of PCHD without above-mentioned complications were divided into Group C. Group B and C was treated with BSPY and hormone replacement therapy respectively, and the drugs for hypolipidemics were withdrawn 1 month before the study. All the patients were observed for 3.5 months, with their blood levels of estradiol (E2) and lipids determined before and after treatment.</p><p><b>RESULTS</b>Before treatment, the level of E2 in the two treated groups was lower than that in the normal group significantly (P < 0.01), and the parameters of blood lipids were abnormal in them. These abnormalities were improved after treatment significantly (P < 0.05 or P < 0.01). The level of E2 raised significantly (P < 0.01) after treatment in patients of Group C, with withdrawal vaginal bleeding presented in 90% of less than 56 years in age. In the Group B after treatment, level of E2 showed a slight rising and withdrawal vaginal bleeding was not found but with improvement of symptoms and signs better than that in Group C.</p><p><b>CONCLUSION</b>Using BSPY in treating PCHD displayed significant adjustment on disturbance of blood lipid spectrum and improvement on clinical manifestations. As compared with the therapeutic effect of hormone replacement therapy, the risk of carcinogenesis caused by endometrial hyperplasia could be avoided because the blood level of E2 is only slightly increased by BSPY.</p>